Abstract
In this paper we first give a brief overview of three-dimensional specimen recording and display using confocal microscopy and digital image processing. We then concentrate on some practical aspects of using this technique: We investigate how the specimen refractive index and detector integration time will influence image resolution and describe an interactive user interface to the computer environment.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Blombäck B, Carlsson K, Hessel B, Liljeborg A, Åslund N (1989) Native fibrin gel networks observed by 3D microscopy, permeation and turbidity. Biochimica et Biophysica Acta 997:96–110
Brakenhoff GJ, van der Voort HTM, van Spronsen EA, Linnemans WAM, Nanninga N (1985) Three-dimensional chromatin distribution in neuroblastoma nuclei shown by confocal scanning laser microscopy. Nature 317:748–749
Brodin L, Ericsson M, Mossberg K, Hökfelt T, Ohta Y, Grillner S (1988) Three-dimensional reconstruction of transmitter-identified central neurons by “en bloc” immunofluorescence histochemistry and confocal scanning microscopy. Experimental Brain Research 73:441–446
Carlsson K (1990a) Scanning and detection techniques used in a confocal scanning laser microscope. Journal of Microscopy 157:21–27
Carlsson K (1990b) 3D representation of microscopic specimens using confocal microscopy and digital image processing. Proceedings of SPIE 1245:68–80
Carlsson K, Åslund N (1987) Confocal imaging for 3-D digital microscopy. Applied Optics 26:3232–3238
Carlsson K, Danielsson PE, Lenz R, Liljeborg A, Majlöf L, Åslund N (1985) Three-dimensional microscopy using a confocal laser scanning microscope. Optics Letters 10:53–55
Carlsson K, Liljeborg A (1989) A confocal laser microscope scanner for digital recording of optical serial sections. Journal of Microscopy 153:171–180
Carlsson K, Wallen P, Brodin L (1989) Three-dimensional imaging of neurons by confocal fluorescence microscopy. Journal of Microscopy 155:15–26
Davidovits P, Egger MD (1971) Scanning laser microscope for biological investigations. Applied Optics 10:1615–1619
Draaijer A, Houpt PM (1987) A real-time confocal laser scanning microscope (CLSM). Proceedings of SPIE 809:85–88
Forsgren PO (1989) Development of methods to perform measurements in two and three dimensions using digital image processing techniques: Applications to limnology, medicine, and nuclear physics. Ph.D. dissertation. Physics IV, The Royal Institute of Technology, Stockholm, Sweden
Fredrikson M, Carlsson K, Franksson O (1988) Confocal scanning laser microscopy, a new technique used in an embryological study of Dactylorhiza maculata (Orchidaceae). Nordic Journal of Botany 8:369–374
Goldstein SR, Hubin T, Rosenthal S, Washburn C (1990) A confocal video-rate laser-beam scanning reflected-light microscope with no moving parts. Journal of Microscopy 157:29–38
Hellmuth T, Seidel P, Siegel A (1988) Spherical aberration in confocal microscopy. Proceedings of SPIE 1028:28–32
Ichioka Y, Kobayashi T, Kitagkwa H, Suzuki T (1985) Digital scanning laser microscope. Applied Optics 24:691–696
Kimura S, Munakata C (1989) Calculation of three-dimensional optical transfer function for a confocal scanning fluorescent microscope. Journal of the Optical Society of America A6:1015–1019
Liljeborg A (1988) Digital position encoding of galvanometer scanner in a laser microscope. Optical Engineering 27:818–822
Marsman HJB, Stricker R, Wijnaendts van Resandt RW, Brakenhoff GJ, Blom P (1983) Mechanical scan system for microscopic applications. Review of Scientific Instruments 54:1047–1052
Minsky M (1961) Microscopy Apparatus, U.S. Patent 3,013,467
Oldmixon EH, Carlsson K, Forsgren PO, Liljeborg A (1988) 3-dimensional reconstructions of lung parenchyma imaged with confocal laser scanning microscope with emphasis on connective tissue cable geometry. Faseb Journal 2:A1270
Petráň M, Hadravský M, Egger MD, Galambos R (1968) Tandem-scanning reflected-light microscope. Journal of the Optical Society of America 58:661–664
Sheppard CJR (1986a) The spatial frequency cut-off in three-dimensional imaging. Optik 72:131–133
Sheppard CJR (1986b) The spatial frequency cut-off in three-dimensional imaging II. Optik 74:128–129
Sheppard CJR (1989) Axial resolution of confocal fluorescence microscopy. Journal of Microcopy 154:237–241
Slomba AF, Wasserman DE, Kaufman GI, Nester JF (1972) A laser flying spot scanner for use in automated fluorescence antibody instrumentation. Journal of the Association for the Advancement of Medical Instrumentation 6:230–234
van der Voort HTM, Brakenhoff GJ, Janssen CGAM, Valkenburg JAC, Nanninga N (1987) Confocal scanning laser fluorescence and reflection microscopy: Measurements of the 3-D image formation and applications in biology. Proceedings of SPIE 809:138–143
Wallén P, Carlsson K, Liljeborg A, Grillner S (1988) Three-dimensional reconstruction of neurons in the lamprey spinal cord in whole-mount, using a confocal laser scanning microscope. Journal of Neuroscience Methods 24:91–100
Wilke V, Gödecke U, Seidel P (1983) Laser-scan-mikroskop. Laser & Optoelektronik 2:93–101
Wilson T, Carlini AR (1987) Size of the detector in confocal imaging systems. Optics Letters 12:227–229
Wilson T, Carlini AR (1989) The effect of aberrations on the axial response of confocal imaging systems. Journal of Microscopy 154:243–256
Wilson T, Sheppard CJR (1984) Theory and practice of scanning optical microscopy. Academic Press, London
Xiao GQ, Kino GS (1987) A real-time confocal scanning optical microscope. Proceedings of SPIE 809:107–113
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Carlsson, K., Lundahl, P. Three-dimensional specimen recording and interactive display using confocal laser microscopy and digital image processing. Machine Vis. Apps. 4, 215–225 (1991). https://doi.org/10.1007/BF01815298
Issue Date:
DOI: https://doi.org/10.1007/BF01815298